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目的观察脑室管膜下区(SVZ)神经干细胞上N-甲基-D-天冬氨酸受体亚单位1(NR1)的超微分布。方法成年大鼠灌注固定,冠状面振动切片,片厚150μm。取含SVZ区域的脑片,行巢蛋白(nestin)和NR1双标免疫组织化学染色。其中,巢蛋白为辣根过氧化物酶标记,NR1为免疫金-银法标记。取1.5mm×1.0mm大小的SVZ双标阳性组织,常规电镜制样、观察、拍照。结果电镜下观察到标记NR1的纳米金-银标颗粒电子密度高,呈颗粒状,分布于细胞膜上,偶见于胞质中;巢蛋白免疫阳性产物呈絮状,分布于胞质中。结论包埋前免疫电镜双重标记技术对组织的抗原性保存较好,免疫金颗粒对抗原定位较精确,有利于研究两种递质在同一个细胞或终末内的共存或分析递质与其相应受体之间的联系,特别是对于中枢神经系统的研究有独到之处。
Objective To observe the ultrastructure of N-methyl-D-aspartate receptor subunit 1 (NR1) in subventricular zone (SVZ) neural stem cells. Methods Adult rats were perfused and fixed with coronal plane vibrational slices with a thickness of 150μm. SVZ-containing brain slices, nestin and NR1 double-stained immunohistochemical staining were used. Among them, Nestin is a horseradish peroxidase marker, and NR1 is an immunogold-silver method marker. Take 1.5mm × 1.0mm size SVZ double-positive tissue, conventional electron microscopy sample, observation, taking pictures. Results Electron microscopy showed that the gold nanoparticles labeled with NR1 had high electron density and granular distribution on the cell membrane, occasionally in the cytoplasm. Nestin immunopositive products were flocculent and distributed in the cytoplasm. Conclusions Immunoelectron microscopy double labeling technique can preserve the antigenicity of the tissue well, and the immunogold particles locate the antigen more precisely, which is helpful for the coexistence or analysis of the two neurotransmitters in the same cell or terminal. The relationship between receptors, especially for the study of the central nervous system unique.